乳酸菌质粒提取方法的建立

Development of A Method for Extraction of Plasmid DNA from Lactic Acid Bacteria

目的建立简便、快速、安全的乳酸菌质粒提取方法。方法将文献报道的乳酸菌质粒提取方法和大肠杆菌质粒提取方法相结合,建立乳酸菌质粒提取方法。通过方法比较、溶菌酶浓度优化和酶切分析,验证该方法的可行性。结果用所建立的方法提取的乳酸菌质粒电泳与酶切图谱与文献报道的提取方法相比均无明显差异。溶菌酶最佳浓度为10mg/ml,避免了毒性物质溴化乙锭的应用,减少了溶菌酶的用量和溶液体积。结论所建立的方法可用于乳酸菌质粒的快速提取,为乳酸菌分子生物学研究奠定了基础。

Objective To develop a simple,rapid and safe method for extracfon of plasmid DNA from lactic acid bacteria.Methods Developa method for extraction of plasmid DNA from lactic acid bacteria by combining the method for extraction of plasmid DNA from E. coli and the method reported. Compare the electrophoretic profiles and restriction analysis results of plasmid DNAs extracted by various methods and optimize the lysozyme concentration to evaluate the feasibility of developed method. Results The electropberetic profile and restriction analysis result of plasmid DNA extracted by the developed method showed no significant difference with those by the method reported. The optimal lysozyme concentration was 10 mg/ml. In addition, by the developed method, the usage toxic ethidium bromide was avoid, and beth the dosage of lysozyme and volume of solution decreased. Conclusion The developed method may be used for the rapid extraction of plasmid DNA from lactic acid bacteria, which laid a fotmdation of molecular biological study on lactic acid bacteria.